Special emphasis has been placed upon development of techniques to improve existing methods for rapid viral diagnosis. The use of a short term tissue culture technique (24 hours) followed by staining of the cells using an anti herpes antibody linked to biotin with the fluoroscein labeled avidin conjugate is a highly efficient system for detecting herpes antigen and is much quicker than detection by conventional tissue culture methods. The correlation of this method with conventional tissue culture was 100%. Changing the parameters of the staining techniques did not help reduce the culture time needed. Several techniques were examined in an attempt to identify the viral antigen directly in the specimen without culture in tissue culture. Staining of the specimen with the biotin-avidin fluoroscein technique was not highly successful. An enzyme-linked immunosorbent assay (ELISA) was developed. Briefly, anti-herpes antibody was attached to the surface of wells in a 96 well plate; plates of the Immulon II or Costar brand were the best. The clinical specimen was incubated and rewashed and anti-herpes antibody linked to biotin was added. After incubation and washing and alkaline phosphatase avidin conjugate was added. After incubation and washing the appropriate substrate was added and the color read. This method has excellent sensitivity (95.6%) and specificity (9l.4%) when compared to conventional tissue culture techniques. The results of this test were obtained within 41/2 hours after initiation of the test procedure. Infectious particles were not needed for the test so that samples not handled correctly for infectivity studies could still be examined for presence of antigen. Routine contamination of tissue cultures for experimental virus studies for mycoplasma contamination continues.